Field Populations Of Flies Research Articles

Single and Combined Mutations of Acetylcholinesterase Gene Giving Resistance to Pirimiphos-Methyl in Musca domestica Slaughterhouse Populations

The house fly Musca domestica L. (Diptera: Muscidae) is a worldwide medical and veterinary pest, causing great economic losses. Organophosphate insecticides have been widely used to control house fly populations. The main objectives of the present study were to evaluate the resistance levels of M. domestica slaughterhouse populations, collected from Riyadh, Jeddah, and Taif, against the organophosphate insecticide pirimiphos-methyl and investigate the genetic mutations of the Ace gene associated with pirimiphos-methyl resistance. The obtained data showed that there were significant differences among pirimiphos-methyl LC50 values of the studied populations, where the highest LC50 was recorded for the Riyadh population (8.44 mM), followed by Jeddah and Taif populations (2.45 mM and 1.63 mM, respectively). Seven nonsynonymous SNPs were detected in the studied house flies. The Ile239Val and Glu243Lys mutations are reported for the first time, whereas Val260Leu, Ala316Ser, Gly342Ala, Gly342Val, and Phe407Tyr were previously reported in M. domestica field populations from other countries. Considering three mutations associated with insecticide resistance, at amino acid positions 260, 342, and 407 of acetylcholinesterase polypeptide, 17 combinations were recovered in this study. Three out of these seventeen combinations were frequently found both worldwide and in the three Saudi house fly field populations, as well as their pirimiphos-methyl-surviving flies. Overall, the single and combined Ace mutations are apparently associated with pirimiphos-methyl resistance, and the obtained data can be useful in managing house fly field populations in Saudi Arabia.

Concentration Dependent Feeding on Imidacloprid by Behaviorally Resistant House Flies, Musca domestica L. (Diptera: Muscidae).

The house fly (Musca domestica L.) is a cosmopolitan and synanthropic pest fly commonly associated with confined animal facilities, known to mechanically vector numerous disease-causing pathogens. Control of adult house flies often relies on insecticides formulated into insecticidal baits, though many baits have failed due to insecticide resistance. House fly resistance to imidacloprid, the most widely used neonicotinoid insecticide available for fly control, has evolved through physiological and behavioral mechanisms in field populations. Behavioral resistance to imidacloprid was documented in field populations of flies from southern California dairies. Lab colonies of these flies were established and behavioral resistance to imidacloprid was selected over several generations. The current study examined the ability of these lab-selected flies to feed on varying concentrations of imidacloprid formulated in sucrose, and if these flies would demonstrate a feeding preference for different concentrations of imidacloprid when exposed in bioassays. Behaviorally resistant flies preferred to feed on untreated sucrose as opposed to treated sucrose at concentrations greater than 25 µg/g imidacloprid when provided sucrose treated with and without imidacloprid. When provisioned with only sucrose treated with a low and high imidacloprid concentration, flies fed on the low concentrations (≤100 µg/g) imidacloprid but reduced feeding on either treatment when concentrations were >100 µg/g imidacloprid. The current study extends the body of knowledge on house fly behavioral resistance to imidacloprid, which could provide insights into future failures of granular fly baits.

Sequence and transcript expression of the super-kdr locus of the horn fly, Haematobia irritans.

In horn flies, Haematobia irritans irritans (Diptera: Muscidae) (Linnaeus, 1758), target site resistance to pyrethroids can be diagnosed by an allele-specific PCR that genotypes individual flies at both the super-kdr (skdr) and the knock down resistance (kdr) associated loci. When this technique uses genomic DNA as template, modifications, such as alternative RNA splicing and RNA editing are not specifically detected. Alternative splicing at the skdr locus has been reported in Dipterans; thus, the genomic DNA-based allele-specific PCR may not accurately reflect the frequency of the skdr mutation in horn fly field populations. To investigate if alternative splicing occurs at the skdr locus of horn flies, genomic DNA and cDNA sequences isolated from two wild populations and two laboratory-reared colonies with varying degrees of pyrethroid resistance were compared. There was no indication of alternative splicing at the super-kdr locus neither in the wild populations nor in the laboratory-reared colonies.

Impacts of long-term insecticide treatment regimes on skdr and kdr pyrethroid resistance alleles in horn fly field populations

We evaluated the effects of four different 6-year duration control strategies on the resistance levels and frequency of the pyrethroid target site resistance alleles, superkdr (skdr) and kdr, at four field populations of Haematobia irritans irritans (Linnaeus, 1758) (Diptera: Muscidae) in Louisiana, USA. Consecutive use of pyrethroid ear tags for 6years caused a significant increase in the resistance ratio to pyrethroids as well as the frequencies of both skdr and kdr resistance alleles. After 3years of consecutive use of pyrethroid ear tags, followed by 1year with no treatment, and followed by 2years with organophosphate ear tags, the resistance ratio for pyrethroid was not significantly affected, the %R-skdr significantly dropped while the %R-kdr allele remained relatively high and stable. Similar results were observed when pyrethroid ear tags were used for three consecutive years, followed by 1year with no treatment, and followed by 2years with endosulfan ear tags; however, this treatment resulted in a slight increase in the resistance ratio for pyrethroids. In a mosaic, the resistance ratio for pyrethroids showed a 2.5-fold increase but the skdr-kdr genetic profiles did not change, as the %R alleles (skdr and kdr) remained low and stable through the 6years. Lack of exposure to pyrethroid insecticides for 3years significantly affected the skdr mutation but not the kdr mutation, preventing re-establishment of susceptibility to pyrethroids. SS-SR (skdr-kdr) individuals were responsible for the maintenance of the kdr mutation in two of the populations studied, and fitness cost seems to strongly affect the SR-RR genotype. None of the four treatment regimens evaluated in the study had satisfactory results for the management of kdr resistance alleles.

Mortality and knockdown effects of imidacloprid and methomyl in house fly (Musca domestica L., Diptera: Muscidae) populations

Mortality and knockdown effects of imidacloprid and methomyl in house fly (Musca domestica L., Diptera: Muscidae) populations

Susceptibility of Spinosad in Musca domestica (Diptera: Muscidae) Field Populations

The toxicity of spinosad was determined in one susceptible and five insecticide-resistant laboratory strains of house fly, Musca domestica L. Spinosad was relatively slow-acting, but highly toxic to house flies. In a feeding bioassay, spinosad LC50 at 72 h was 0.51 microg of spinosad per gram of sugar, making it 6.3- and 3.5-fold more toxic to house flies compared with azamethiphos and methomyl, respectively. In topical application bioassay, the LD50 at 48 h of spinosad in susceptible house flies was 40 ng per 20 mg of house fly, making spinosad less toxic than the pyrethroid bioresmethrin synergized by piperonyl butoxide and the organophosphate dimethoate. The insecticide-resistant laboratory strains had resistance factors to spinosad at LC50 in feeding bioassay from 1.5 to 5.5 and at LD50 in topical application bioassay from 2.5 to 4.7, indicating that in house fly cross-resistance to the major insecticide classes will not initially be of major concern for the use of spinosad for house fly control. The toxicity of spinosad was also evaluated against 31 field populations of house flies collected from livestock farms across Denmark. The field populations were 2.2- to 7.5-fold resistant to spinosad at 72 h in feeding bioassay, but based on steep slopes in the bioassay and the limited variation of spinosad toxicity against the various field populations, we consider the field populations to be spinosad-susceptible. We propose a diagnostic dose of 12 microg of spinosad per gram of sugar in feeding bioassay with impregnated sugar for determination of resistant house flies, which is 10x the LC95 of the susceptible strain WHO and approximately = 2x the LD95 of the field populations. Spinosad showed no substantial cross-resistance to the pyrethroid bioresmethrin synergized by piperonyl butoxide, the anticholinesterases dimethoate, azamethiphos, methomyl, and spinosad in house fly field populations.

Frequency of <I>kdr</I> Gene in House Fly Field Populations: Correlation of Pyrethroid Resistance and <I>kdr</I> Frequency

Frequency of <I>kdr</I> Gene in House Fly Field Populations: Correlation of Pyrethroid Resistance and <I>kdr</I> Frequency

Frequency of kdr Gene in House Fly Field Populations: Correlation of Pyrethroid Resistance and kdr Frequency

The frequency of the L1014 F kdr mutation was determined in 14 field populations of house flies, Musca domestica L., with resistance factors at LD50 for pyrethrin/piperonyl butoxide and bioresmethrin/piperonyl butoxide from 4 to 29 and 2 to 98, respectively. A polymerase chain reaction test for identifying kdr homo- or heterozygote house flies was used to determine the frequency of kdr. The L1014 F allele was found in all populations tested. The frequency of kdr in the field populations was high and varied from 0.46 to 0.99. Eleven of the populations were in Hardy-Weinberg equilibrium, whereas two strains had higher number of heterozygotes than expected, indicating a possible heterozygote advantage. The frequency of kdr was strongly correlated with the reduced mortality observed in the bioassays with pyrethrum and bioresmethrin synergized by piperonyl butoxide. This indicates that kdr is a major mechanism for pyrethroid resistance in these field populations. Five field populations had resistance factors >25 and >10 for bioresmethrin/piperonyl butoxide and pyrethrin/piperonyl butoxide, respectively. The frequencies of kdr in these five populations varied from 0.89 to 0.99. The frequencies of kdr in the field populations showing no or a low level of resistance had frequencies of kdr from 0.46 to 0.75, which indicates that the L1014 F kdr allele is a fully recessive genetic trait in house flies. We have shown that the molecular diagnostic PASA method to determine the resistance phenotypes and the frequency of kdr is a powerful tool, which could be used to get information to make recommendations about pest and resistance management.

Susceptibility of Spinosad in <I>Musca domestica</I> (Diptera: Muscidae) Field Populations

The toxicity of spinosad was determined in one susceptible and five insecticide-resistant laboratory strains of house fly, Musca domestica L. Spinosad was relatively slow-acting, but highly toxic to house flies. In a feeding bioassay, spinosad LC50 at 72 h was 0.51 microg of spinosad per gram of sugar, making it 6.3- and 3.5-fold more toxic to house flies compared with azamethiphos and methomyl, respectively. In topical application bioassay, the LD50 at 48 h of spinosad in susceptible house flies was 40 ng per 20 mg of house fly, making spinosad less toxic than the pyrethroid bioresmethrin synergized by piperonyl butoxide and the organophosphate dimethoate. The insecticide-resistant laboratory strains had resistance factors to spinosad at LC50 in feeding bioassay from 1.5 to 5.5 and at LD50 in topical application bioassay from 2.5 to 4.7, indicating that in house fly cross-resistance to the major insecticide classes will not initially be of major concern for the use of spinosad for house fly control. The toxicity of spinosad was also evaluated against 31 field populations of house flies collected from livestock farms across Denmark. The field populations were 2.2- to 7.5-fold resistant to spinosad at 72 h in feeding bioassay, but based on steep slopes in the bioassay and the limited variation of spinosad toxicity against the various field populations, we consider the field populations to be spinosad-susceptible. We propose a diagnostic dose of 12 microg of spinosad per gram of sugar in feeding bioassay with impregnated sugar for determination of resistant house flies, which is 10x the LC95 of the susceptible strain WHO and approximately = 2x the LD95 of the field populations. Spinosad showed no substantial cross-resistance to the pyrethroid bioresmethrin synergized by piperonyl butoxide, the anticholinesterases dimethoate, azamethiphos, methomyl, and spinosad in house fly field populations.

Possible Mechanism of Diazinon Negative Cross-Resistance in Pyrethroid-Resistant Horn Flies (Diptera: Muscidae)

The underlying mechanism responsible for enhanced toxicity of the organophosphate diazinon in eight pyrethroid-resistant horn fly, Haematohia irritans (L.), field populations was investigated. In vitro enzyme assays and in vivo pyrethroid synergist studies showed increased activity of the mixed function oxidase system (measured as O -demethylase activity) in five of the eight populations with, in most instances, a subsequent increase in diazinon metabolism. We suspect that enhanced toxicity of diazinon (or negative cross-resistance), as observed in pyrethroid-resistant horn fly field populations, may result from in vivo accumulation of the toxic metabolite diazoxon.

Stable Fly (Diptera: Muscidae) Migration in Northwest Florida

Migration of the stable fly, Stomoxys calcitrans (L.), in northwest Florida and its relationship to weather patterns were investigated. Field populations of flies were marked with fluorescent dust using Attractant Self-Marking Devices and captured on Williams traps. A new flight range of 225 km was established for the species. This was the first migration study performed with wild stable flies where flies were not handled until they were captured on Williams traps. By the use of habitat-specific mites phoretically attached to stable flies, physiological age-grading, and sex ratios of flies captured on Williams traps, beach populations of stable flieswere shown to be migratory.